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TABLE 1 (Continued) List of fertility associated proteins highly abundant in high fertile bulls along with their functions and log fold changes

Table 1 (Continued) List of fertility associated proteins highly abundant in high fertile bulls along with their functions and log fold changes

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FIGURE 1 Selection of high and low fertile buffalo bulls based on conception rate (CR). (A) Five bulls with highest CRs were considered as high fertile while five bulls with lowest CRs were considered as low fertile. (B) The mean CRs of shortlisted high and low fertile bulls were significantly different (jp < 0.0001)
FIGURE 1 Selection of high and low fertile buffalo bulls based on conception rate (CR). (A) Five bulls with highest CRs were considered as high fertile while five bulls with lowest CRs were considered as low fertile. (B) The mean CRs of shortlisted high and low fertile bulls were significantly different (jp < 0.0001)
FIGURE 2 Sperm viability and acrosome integrity tests in HF and LF spermatozoa. (A) Cell viability assay through membrane integrity employing CFDA with Pl was use to categorized sperms as live, dead and moribund in high fertile (a, b, c) and low fertile (d, e, f) groups. (B) Histogram plots showing percentage of different categories of sperms observed in the viability test. (C) Acrosome integrity test employing FITC- PNA along with PI was used to categorize the spermatozoa as acrosome intact or acrosome-reacted in high fertile (g, h, i) and low fertile (j, k, |) spermatozoa (original magnification 60X). (D) Histogram plots showing percentage of different categories of sperms observed in the acrosome integrity test.
FIGURE 2 Sperm viability and acrosome integrity tests in HF and LF spermatozoa. (A) Cell viability assay through membrane integrity employing CFDA with Pl was use to categorized sperms as live, dead and moribund in high fertile (a, b, c) and low fertile (d, e, f) groups. (B) Histogram plots showing percentage of different categories of sperms observed in the viability test. (C) Acrosome integrity test employing FITC- PNA along with PI was used to categorize the spermatozoa as acrosome intact or acrosome-reacted in high fertile (g, h, i) and low fertile (j, k, |) spermatozoa (original magnification 60X). (D) Histogram plots showing percentage of different categories of sperms observed in the acrosome integrity test.
FIGURE 3 Venn diagrams representing the total identified proteins in HF and LF spermatozoa. The protein analytics tool Proteome Discoverer (v2.2) was used and 1,002 proteins were found common in both HF and LF sperms while 288 and 95 unique proteins were identified in HF and LF respectively wit isoforms (p < 0.05, FDR < 0.01). Some of the proteins were known in nature, followed by predicted, uncharacterized (UC), and LOC proteins.
FIGURE 3 Venn diagrams representing the total identified proteins in HF and LF spermatozoa. The protein analytics tool Proteome Discoverer (v2.2) was used and 1,002 proteins were found common in both HF and LF sperms while 288 and 95 unique proteins were identified in HF and LF respectively wit isoforms (p < 0.05, FDR < 0.01). Some of the proteins were known in nature, followed by predicted, uncharacterized (UC), and LOC proteins.
FIGURE 4 Representation of differentially abundant proteins (DAPs) in high and low fertile spermatozoa. (A) Heat map showing differentially abundant proteins (DAPs) among the replicates of HF and LF groups. Intense blue colour represents high abundance of proteins while white colour represents low abundance of proteins. (B) Volcano plot of all DAPs identified in the proteomics data determine by log fold change vs. —log10 p-value. Red points: DAPs that were significantly high abundant in high fertile bull (fold change >2; p < 0.05). Blue points: DAPs that were significantly low abundant in high fertile bull (fold change <0.5; p < 0.05). Grey points: DAPs that showed neutral abundance. Volcano plot showing the significantly abundant proteins determine by log fold change (log fold) vs. -log10 p-value. (C) PCA plot representing the level of variances between the replicates of HF and LF samples.
FIGURE 4 Representation of differentially abundant proteins (DAPs) in high and low fertile spermatozoa. (A) Heat map showing differentially abundant proteins (DAPs) among the replicates of HF and LF groups. Intense blue colour represents high abundance of proteins while white colour represents low abundance of proteins. (B) Volcano plot of all DAPs identified in the proteomics data determine by log fold change vs. —log10 p-value. Red points: DAPs that were significantly high abundant in high fertile bull (fold change >2; p < 0.05). Blue points: DAPs that were significantly low abundant in high fertile bull (fold change <0.5; p < 0.05). Grey points: DAPs that showed neutral abundance. Volcano plot showing the significantly abundant proteins determine by log fold change (log fold) vs. -log10 p-value. (C) PCA plot representing the level of variances between the replicates of HF and LF samples.
TABLE 1 List of fertility associated proteins highly abundant in high fertile bulls along with their functions and log fold changes.
TABLE 1 List of fertility associated proteins highly abundant in high fertile bulls along with their functions and log fold changes.
(Continued on following page) TABLE 1 (Continued) List of fertility associated proteins highly abundant in high fertile bulls along with their functions and log fold changes
(Continued on following page) TABLE 1 (Continued) List of fertility associated proteins highly abundant in high fertile bulls along with their functions and log fold changes
TABLE 2 List of fertility associated proteins low abundant in high fertile bulls along with their functions and log fold changes.
TABLE 2 List of fertility associated proteins low abundant in high fertile bulls along with their functions and log fold changes.
Pie charts representing top 10 Gene Ontology (GO) terms with respect to Biological Process (BP), Molecular Functions (MF) and Cellular Component (CC) (A) high abundant protein in High fertile (B) low abundant protein in High fertile spermatozoa.
Pie charts representing top 10 Gene Ontology (GO) terms with respect to Biological Process (BP), Molecular Functions (MF) and Cellular Component (CC) (A) high abundant protein in High fertile (B) low abundant protein in High fertile spermatozoa.
Validation of differential abundance of AKAP3, Sp17, and DLD using western blot. (A) The blot image showing relative abundance of AKAP3, Sp17, and DLD in HF and LF samples taking B-actin as calibrator. (B) Histograms representing the relative intensities of AKAP3, Sp17, and DLD among the HF and LF samples that were calculated using the blot images in the ImageJ software.
Validation of differential abundance of AKAP3, Sp17, and DLD using western blot. (A) The blot image showing relative abundance of AKAP3, Sp17, and DLD in HF and LF samples taking B-actin as calibrator. (B) Histograms representing the relative intensities of AKAP3, Sp17, and DLD among the HF and LF samples that were calculated using the blot images in the ImageJ software.
FIGURE 7 The cellular localization of (A) AKAP3, (B) DLD, and (C) Sp17 in HF and LF spermatozoa using FITC-labelled antibodies. Nuclear staining was carri¢ out using DAPI. The images were captured at x600 magnification.
FIGURE 7 The cellular localization of (A) AKAP3, (B) DLD, and (C) Sp17 in HF and LF spermatozoa using FITC-labelled antibodies. Nuclear staining was carri¢ out using DAPI. The images were captured at x600 magnification.
The Violin plots representing the relative abundance of various DAPs between the HF and LF groups. In the plots, y-axes represent log2 intensities o the respective DAPs. (A) high abundant protein in High fertile (B) low abundant protein in High fertile spermatozoa.
The Violin plots representing the relative abundance of various DAPs between the HF and LF groups. In the plots, y-axes represent log2 intensities o the respective DAPs. (A) high abundant protein in High fertile (B) low abundant protein in High fertile spermatozoa.
Schematic representation of fertility associated high and low abundant proteins in HF spermatozoa.
Schematic representation of fertility associated high and low abundant proteins in HF spermatozoa.
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AndrologyBiologyArtificial inseminationSperm Motility
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